白藜芦醇对氧糖剥夺/再灌注损伤的PC12细胞的保护作用及其作用机制

Protective Effect of Resveratrol on the Injury of PC12 Cells Induced by OGD/R and Its Mechanisms

目的:探讨白藜芦醇对氧糖剥夺/再灌注(OGD/R)损伤的PC12细胞的保护作用及其机制。方法:体外培养PC12细胞,分为对照组,白藜芦醇组,OGD/R组及OGD/R+白藜芦醇组。以改良的噻唑蓝法测定细胞活性,采用Annexin V-FITC/PI双染法检测细胞的凋亡率,用双氢罗丹明(DHR)检测细胞内活性氧簇(ROS)的水平,采用蛋白印迹法(western blot)分析SIRT1的蛋白表达情况。结果:与对照组相比,经过OGD/R损伤后,细胞活力显著降低。而在OGD/R的同时给予10μmol/L的白藜芦醇处理,可以明显提高细胞活力。流式细胞仪检测发现,10μmol/L的白藜芦醇可以显著地减少OGD/R引起的细胞凋亡,抑制细胞内的ROS产生。western blot的结果提示,与对照组比较,白藜芦醇可提高SIRT1的蛋白表达水平。结论:白藜芦醇可以通过抑制ROS的产生和上调SIRT1的表达等机制而发挥其对抗氧糖剥夺/再灌注损伤的神经保护性作用。

Objective： To investigate the protective effect of resveratrol （RSV） on the injury of PC12 cells induced by oxygen glucose deprivation/reperfusion （OGD/R） and its mechanisms. Methods： PC12 cells cultured in vitro were divided into control group, RSV group, OGD/R group and OGD/R＋RSV group. The cell viability was assessed by the modified MTT method. Annexin V-FITC/PI double staining was used to detect apoptosis rate. DHR123 was used to measure cellular ROS levels. Western blot was used to study the expression level of SIRT1. Remits： Compared to the normal control group, the cell viability decreased significantly after OGD/R and giving 10μmol/L RSV during OGD/R could significantly increase cell viability. Based on the results of flow cytometry, we found 10 μmol/L RSV markedly inhibited OGD/R-induced apoptosis and reduced intracellular ROS generation. Result of Western blot showed that RSV could increase the expression of SIRT1 compared with control group. Conclusion： RSV was found to have a neuroprotective effect against OGD/R-induced injury by attenuating ROS production and up-regulating the expression level of SIRT1.